Safety

Risks to the safety and health of team members, or other people working in the lab

The use of laboratory coats and latex gloves will be used when handling E.coli to avoid contamination of the cultures and thus avoid spoiling the cultures. However, it is conceivable that latex gloves may pose an allergy hazard or risk of choking (if swallowed). Use of Bunsen burners within the laboratory to preserve a sterile environment may naturally also pose a fire hazard if knocked over and the laboratory safety manual recommends that people with longer hair have it tied back to avoid risks of having it burnt. The use of ethidium bromide in the lab poses a risk of contamination from the carcinogen, to avoid this a separate area of the lab will be dedicated to this and a separate waste area will be provided as well as gloves being changed each time an ethidium bromide contaminated item is touched.

Although, outside of human error, there is little risk posed by the biological materials used and no further considerations will be made regarding the use of laboratory equipment than is already stated by our in-house manuals or any applicable regulations.

Risks to the safety and health of the general public (if any biological materials escaped from your lab)

E.coli K-12 is a non-pathogenic strain that is used in many laboratories and there is naturally no risk to the general public if it were to breakout from the laboratory. The Huh7 cell line is an immortal tumorigenic human cell line that also has no risks to the general public unless it were somehow injected into a immunocompromised individual. Our RNA construct has no danger outside the laboratory as we are not using a transfection vector outside of a company kit and thus the raw construct will be unable to transfect human cells and is not infectious. Therefore, no additional precautions will be taken than is already stated by the in-house manual regulations or any applicable regulations.

Risks to the environment (from waste disposal, or from materials escaping from your lab)

Biological materials are typically disposed of within specified laboratory bins that are subsequently sterilised to prevent the material from being released into the environment via waste. None of our materials are infectious and it is inconceivable how the materials can escape from the laboratory outside of human error. Therefore, no further precautions will be taken than is already stated by the in-house manual regulations or any applicable regulations. We understand that the use and modification of an organism by introduction of genetic material creates a transgenic organism, which is to be confined solely to a lab and no other place. Disposal of ethidium bromide waste products (e.g. gels, contaminated buffer, gloves and pipette tips are handled with a specialized container - contaminated buffer is taken and dealt with by senior technicians).

Risks to security through malicious mis-use by individuals, groups, or countries

We are not aware of any organisations, groups or individuals that are aware of our project unless there are information security compromises on the part of iGEM. Furthermore, we do not foresee any risk to security as a result of our work and there are no applications for misuse for our project.

What measures are you taking to reduce these risks? (For example: safe lab practices, choices of which organisms to use.)

Therefore, no further precautions will be taken than is already stated by the in-house manual regulations or any applicable regulations as the experimental protocols are highly standardised and we have deliberately chosen biological materials (e.g. E.coli, cell lines) that pose no risk to normal individuals working the laboratory.

What new risks might arise from your project's growth?

Whilst all the biological materials used will be of the lowest possible risk group, we do not envision any risks as a result of the project as we are designing an alternative platform of an existing treatment for disease. The university is a well-secured environment and access to the laboratories, buildings etc require card access. Therefore, apart from in-house sabotage from parties of conflicting interest (of which we have none), we do not imagine a security risk to members of the university should knowledge of our project (that will be already made available to the jamboree and thus, available to the public) be made any more available than is currently. As the UK is generally a safe and well-governed country and our project is not of a controversial nature, we do not perceive any possible acts of misuse (e.g. terrorism) possible within the purview of the project.

Does your project currently include any design features to reduce risks?

Our in-vivo RNA construct will have an aptazyme kill switch that responds to theophyline, thus, there are chemical constraints in place to destroy the construct should we wish. We also have a self regulatory MS2 box protein which will help to regulate the replicon. For a full consideration of safety and risks, please see our human practices section in the future.

We have chosen and planned out our project with each person being assigned a particular role in the group in order to get started with every aspect of the scheme.

We uploaded the spreadsheet here .